SRI International and a group of collaborators propose to further develop the EcoCyc database (DB). The DB is accessible to scientists through the World Wide Web via a user-friendly graphical interface. EcoCyc describes the gene and metabolic pathways to E. coli, and its transporters and transcriptional regulation machinery. It serves as a general reference source for E. coli, and is particularly well suited for use in the analysis of microbial genomes. This project will create new DB modules for several pathogenic E. coli strains whose complete DNA sequence will soon be in the public domain. This addition will facilitate comparative analyses of these strains. EcoCyc is a unique resource for analysis of large gene-expression datasets because it couples an extensive collection of data on transcriptional regulatory mechanisms with a unique program for displaying expression datasets on a full pathway map of the cell. When used as a basis for analyzing newly sequenced microbial genomes, EcoCyc contributes to the identification within those genomes of metabolic pathways that may serve as targets for drug design. Pharmaceutical companies are seeking targets within metabolic pathways as one strategy for developing next-generation drugs against antibiotic-resistant bacteria. EcoCyc is also used for teaching biochemistry by colleges and Universities. SRI further proposes to develop new bioinformatics methods for comparative genomics, and for predicting nutritional requirements and essential genes of an organism form its metabolic network.